The proposed research will investigate the possibility of underlying common mechanisms between alcohol and opiates. Recently it has been shown that endorphins are derived from the same precursor molecule and are released together into the blood stream in response to stress. Severe disruptions of ACTH levels often accompany the clinical syndrome of alcoholism. Some studies suggest that alcohol dependence may be mediated via endogenous opiate peptides. It has long been known that alocholism has a familial pattern of incidence, and recent studies have rigorously demonstrated a genetic component in the disorder. In the proposed research, we will systematically study the in vitro effect of ethanol on endorphin and ACTH regulation. Beginning with mouse pituitary tumor cells in culture, we will characterize the effects of acute and chronic ethanol on biosynthesis, processing and release of endorphins and ACTH by: labelling the tumor cells with radioactive amino acids and sugars; treating the cell with ethanol in the presence and absence of known physiological regulators (CRF and glucocorticoids); and finally, immunoprecipitating the various forms of ACTH and endorphin with purified antisera to both hormones. Separation of these forms by SDS/PAGE should provide evidence for or against any effect of ethanol to change biosynthesis, processing and release of ACTH and endorphins. In the second phase, we will separately culture anterior and intermediate-posterior pituitary cells from mice that have been made dependent on ethanol by being treated chronically using a liquid diet technique. These cell cultures will be systematically investigated as described above. Finally, we will extend the investigations to include several inbred strains of mouse. We have characterized 20 inbred mouse strains for sensitivity to, tolerance to and physical dependence on ethanol. From these, we will select for study a range of strains that will enable us to assess the possibility that response to ethanol and regulation of ACTH and endorphin activity may be controlled by common genes.